| Vector | lentivirus |
| Construct | scFv-CD3ζ|scFv-CD28-CD3ζ |
| Vector type | self-inactivating lentiviral vector pHIV7 |
| Transgene/Inserted gene | VH and VL gene segments (scFv) of the CD19-specific FMC63 monoclonal antibody (mAb); IgG4 hinge-CH2-CH3; signaling domains CD3ζ (CD19CAR) |
| Regulatory element | EF-1 promoter |
| Viral genome modification | The CD19R:ζ DNA sequence (optimized by GeneArt) was cloned into a self-inactivating lentiviral vector pHIV7 that has an EF-1 promoter. |
| Vector production method | The huEGFRt was synthesized by PCR splice overlap extension to fuse in frame the human granulocyte-macrophage-colony-stimulating factor receptor's leader peptide to domains III and IV and the transmembrane spanning components of huEGFR (base pairs, 1000-2004). This fusion product was then cloned into the epHIV7 vector (in which the cytomegalovirus promoter of pHIV7 was replaced with an EF-1 promoter) along with the CD19CAR and T2A sequences, and the final construct was confirmed by sequence analysis. |
| Additional feature | EGFRt|cytoplasmic domain of the CD3ζ chain|CD28 costimulatory domain|self-cleaving T2A sequence |